菊芋不同部位DNA提取的比较研究

Comparative study on extraction of genomic DNA from different parts of Helianthus tuberosus L.

分别以菊芋的茎段、叶片和块茎为材料,采用改良CTAB法分别提取菊芋3个部位的基因组DNA,用0.8%的琼脂糖凝胶电泳检测DNA质量,分别测定3个部位的DNA在A260和A280下的吸光值,根据A260/A280的值检测DNA的浓度和纯度,并以3个部位提取的DNA为模板进行ISSR扩增。结果表明:采用菊芋叶片提取DNA的产率最高,茎段次之,块茎最低。三个部位提取的DNA纯度均相近,提取叶片获得的DNA浓度最大,为163.2 ng·μL-1;其次是茎段,为137.4 ng·μL-1;块茎获得的DNA浓度最低,为95.5 ng·μL-1。菊芋不同部位提取的DNA模板对ISSR扩增没有明显影响,DNA浓度都能达到扩增的要求。菊芋3个部位提取的DNA质量均较好,可以进行后续的酶切和PCR扩增等实验。

With the stems, young leaves and tubers of Helianthus tuberosus L. as experimental materials, the genome DNA were extracted with modified CTAB method and examined with 0. 8% agarose gel electrophoresis. The absor- bance value of DNA from these parts was measured by ultraviolet spectrophotometer at A260 and A280 and the ratio of A260/A280 was used to determine the DNA purity and concentration. DNA extracted from these parts of Helianthus tu- berosus L. was used as the template for ISSR amplification. The results showed the yield of DNA extracted from Heli- anthus tuberosus L. leaves was the highest, that from the stems was the second, and that from the tubers was the low- est. The purity of DNA extracted from these parts of Helianthus tuberosus L. was similar. The concentration of DNA extracted from Helianthus tuberosus L. leaves was the the largest （ 163.2 ng. μL-1 ） followed by that from stems （137.4 ng μL-1 ）, and that from the tubers was the lowest （95. 5 ng μL-1 ）. The DNA template extracted from different parts of Helianthus tuberosus L. had rio significant effect on ISSR amplification, and DNA concentration could achieve the amplification requirements. The DNA quality extracted from above three parts of Helianthus tu- berosus L. was good and could be used in subsequent expermients such as digestion and PCR amplification.